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目的 探讨HIV - 1反式激活蛋白 (TAT)的蛋白转导结构域 (PTD)介导的BCR/ABL(TATPTD -BCR/ABL)融合蛋白通过血脑屏障的作用。方法 在大肠杆菌中表达PTD -BCR/ABL融合蛋白。将经Ni-NTA树脂亲和层析纯化的融合蛋白 ,经尾静脉注射小鼠体内 ,用免疫组织化学染色法 ,对小鼠脑组织细胞内的PTD -BCR/ABL融合蛋白进行检测。结果 ①表达和纯化了相对分子质量 (Mr)为 2 3× 10 3 的PTD -BCR/ABL融合蛋白 ;②将其经静脉注射到小鼠体内 ,在小鼠脑组织细胞中可检测到PTD -BCR/ABL融合蛋白。结论 PTD可在体内介导较大Mr的融合蛋白通过血脑屏障 ,有望为治疗性的药物通过血脑屏障进入中枢神经系统提供了一种新的方法
Objective To investigate the effect of BTR / ABL (TATPTD-BCR / ABL) fusion protein passing through the blood-brain barrier mediated by the protein transduction domain (PTD) of HIV-1 transactivator protein (TAT) Methods The PTD-BCR / ABL fusion protein was expressed in E. coli. The fusion protein purified by Ni-NTA resin affinity chromatography was injected into the tail vein of mice, and the PTD-BCR / ABL fusion protein in mouse brain cells was detected by immunohistochemical staining. Results ① Expression and purification of PTD - BCR / ABL fusion protein with relative molecular mass (Mr) of 2 3 × 10 3; (2) PTD - BCR / ABL fusion protein was injected intravenously into mice. PTD - BCR / ABL fusion protein. Conclusions PTD can mediate the greater Mr fusion protein crossing the blood-brain barrier in vivo and is expected to provide a new method for therapeutic drug entry into the central nervous system through the blood-brain barrier